CN111732576B - 生物正交激活的时间分辨响应型稀土探针及其制备方法与应用 - Google Patents
生物正交激活的时间分辨响应型稀土探针及其制备方法与应用 Download PDFInfo
- Publication number
- CN111732576B CN111732576B CN201911266381.7A CN201911266381A CN111732576B CN 111732576 B CN111732576 B CN 111732576B CN 201911266381 A CN201911266381 A CN 201911266381A CN 111732576 B CN111732576 B CN 111732576B
- Authority
- CN
- China
- Prior art keywords
- rare earth
- keto
- activated
- amino
- probe
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
- 239000000523 sample Substances 0.000 title claims abstract description 68
- 229910052761 rare earth metal Inorganic materials 0.000 title claims abstract description 60
- 150000002910 rare earth metals Chemical class 0.000 title claims abstract description 57
- 230000004044 response Effects 0.000 title claims abstract description 26
- 238000002360 preparation method Methods 0.000 title claims abstract description 21
- 238000006243 chemical reaction Methods 0.000 claims abstract description 34
- 238000003384 imaging method Methods 0.000 claims abstract description 23
- DPOPAJRDYZGTIR-UHFFFAOYSA-N Tetrazine Chemical group C1=CN=NN=N1 DPOPAJRDYZGTIR-UHFFFAOYSA-N 0.000 claims abstract description 13
- 210000004881 tumor cell Anatomy 0.000 claims abstract description 12
- 241000252212 Danio rerio Species 0.000 claims abstract description 11
- 210000000170 cell membrane Anatomy 0.000 claims abstract description 11
- -1 rare earth ions Chemical class 0.000 claims abstract description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 8
- 230000004913 activation Effects 0.000 claims abstract description 7
- 238000000034 method Methods 0.000 claims abstract description 7
- WVFGZDBVYAGUCQ-UHFFFAOYSA-N 7-amino-2-oxo-1H-quinoline-4-carboxylic acid Chemical compound C1=CC2=C(C=C1N)NC(=O)C=C2C(=O)O WVFGZDBVYAGUCQ-UHFFFAOYSA-N 0.000 claims description 27
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 24
- 239000007787 solid Substances 0.000 claims description 22
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 21
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 16
- ZWKQKWLZKSZYAT-UHFFFAOYSA-N [4-(6-methyl-1,2,4,5-tetrazin-3-yl)phenyl]methanamine Chemical compound N1=NC(C)=NN=C1C1=CC=C(CN)C=C1 ZWKQKWLZKSZYAT-UHFFFAOYSA-N 0.000 claims description 15
- 238000003756 stirring Methods 0.000 claims description 15
- 210000004027 cell Anatomy 0.000 claims description 12
- FALRKNHUBBKYCC-UHFFFAOYSA-N 2-(chloromethyl)pyridine-3-carbonitrile Chemical compound ClCC1=NC=CC=C1C#N FALRKNHUBBKYCC-UHFFFAOYSA-N 0.000 claims description 9
- PIICEJLVQHRZGT-UHFFFAOYSA-N Ethylenediamine Chemical compound NCCN PIICEJLVQHRZGT-UHFFFAOYSA-N 0.000 claims description 9
- 229940014800 succinic anhydride Drugs 0.000 claims description 9
- 230000003213 activating effect Effects 0.000 claims description 8
- ULJUVCOAZNLCJZ-UHFFFAOYSA-K trichloroterbium;hexahydrate Chemical compound O.O.O.O.O.O.[Cl-].[Cl-].[Cl-].[Tb+3] ULJUVCOAZNLCJZ-UHFFFAOYSA-K 0.000 claims description 8
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 7
- 238000001035 drying Methods 0.000 claims description 7
- 239000000047 product Substances 0.000 claims description 7
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 claims description 6
- NQTADLQHYWFPDB-UHFFFAOYSA-N N-Hydroxysuccinimide Chemical compound ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 claims description 6
- 239000002904 solvent Substances 0.000 claims description 6
- 239000002253 acid Substances 0.000 claims description 5
- 230000001376 precipitating effect Effects 0.000 claims description 5
- 239000007795 chemical reaction product Substances 0.000 claims description 4
- 238000010898 silica gel chromatography Methods 0.000 claims description 4
- 206010058467 Lung neoplasm malignant Diseases 0.000 claims description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 2
- 239000003795 chemical substances by application Substances 0.000 claims description 2
- 201000005202 lung cancer Diseases 0.000 claims description 2
- 208000020816 lung neoplasm Diseases 0.000 claims description 2
- 239000002244 precipitate Substances 0.000 claims description 2
- 239000000741 silica gel Substances 0.000 claims description 2
- 229910002027 silica gel Inorganic materials 0.000 claims description 2
- WGQKYBSKWIADBV-UHFFFAOYSA-N benzylamine Chemical compound NCC1=CC=CC=C1 WGQKYBSKWIADBV-UHFFFAOYSA-N 0.000 claims 4
- 239000003153 chemical reaction reagent Substances 0.000 claims 3
- 238000004519 manufacturing process Methods 0.000 claims 2
- 230000001413 cellular effect Effects 0.000 claims 1
- 238000013375 chromatographic separation Methods 0.000 claims 1
- 239000012528 membrane Substances 0.000 claims 1
- 238000002156 mixing Methods 0.000 claims 1
- 230000008859 change Effects 0.000 abstract description 6
- DVKJUZGVRLQSDH-WKWWPMDXSA-N C1=CCCCCCC1.O=C[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO Chemical compound C1=CCCCCCC1.O=C[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO DVKJUZGVRLQSDH-WKWWPMDXSA-N 0.000 abstract description 5
- 238000002866 fluorescence resonance energy transfer Methods 0.000 abstract description 3
- 230000008569 process Effects 0.000 abstract description 3
- 239000000090 biomarker Substances 0.000 abstract description 2
- 238000002165 resonance energy transfer Methods 0.000 abstract description 2
- 238000001095 inductively coupled plasma mass spectrometry Methods 0.000 description 7
- URYYVOIYTNXXBN-OWOJBTEDSA-N trans-cyclooctene Chemical compound C1CCC\C=C\CC1 URYYVOIYTNXXBN-OWOJBTEDSA-N 0.000 description 7
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 6
- 238000012512 characterization method Methods 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- 238000002189 fluorescence spectrum Methods 0.000 description 5
- 102000003886 Glycoproteins Human genes 0.000 description 4
- 108090000288 Glycoproteins Proteins 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- 238000010226 confocal imaging Methods 0.000 description 4
- URYYVOIYTNXXBN-UPHRSURJSA-N cyclooctene Chemical compound C1CCC\C=C/CC1 URYYVOIYTNXXBN-UPHRSURJSA-N 0.000 description 4
- 239000004913 cyclooctene Substances 0.000 description 4
- 238000011534 incubation Methods 0.000 description 4
- 238000009616 inductively coupled plasma Methods 0.000 description 4
- 238000001819 mass spectrum Methods 0.000 description 4
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical class [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 3
- 238000002372 labelling Methods 0.000 description 3
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 206010027476 Metastases Diseases 0.000 description 2
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 2
- 238000000862 absorption spectrum Methods 0.000 description 2
- 238000004624 confocal microscopy Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 238000003745 diagnosis Methods 0.000 description 2
- 239000007850 fluorescent dye Substances 0.000 description 2
- 230000009401 metastasis Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 2
- 235000019796 monopotassium phosphate Nutrition 0.000 description 2
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- NGNBDVOYPDDBFK-UHFFFAOYSA-N 2-[2,4-di(pentan-2-yl)phenoxy]acetyl chloride Chemical class CCCC(C)C1=CC=C(OCC(Cl)=O)C(C(C)CCC)=C1 NGNBDVOYPDDBFK-UHFFFAOYSA-N 0.000 description 1
- 241000251468 Actinopterygii Species 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- JPVYNHNXODAKFH-UHFFFAOYSA-N Cu2+ Chemical compound [Cu+2] JPVYNHNXODAKFH-UHFFFAOYSA-N 0.000 description 1
- 238000005698 Diels-Alder reaction Methods 0.000 description 1
- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical compound [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- QPCDCPDFJACHGM-UHFFFAOYSA-N N,N-bis{2-[bis(carboxymethyl)amino]ethyl}glycine Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(=O)O)CCN(CC(O)=O)CC(O)=O QPCDCPDFJACHGM-UHFFFAOYSA-N 0.000 description 1
- WTBIAPVQQBCLFP-UHFFFAOYSA-N N.N.N.CC(O)=O.CC(O)=O.CC(O)=O.CC(O)=O.CC(O)=O Chemical compound N.N.N.CC(O)=O.CC(O)=O.CC(O)=O.CC(O)=O.CC(O)=O WTBIAPVQQBCLFP-UHFFFAOYSA-N 0.000 description 1
- 238000005481 NMR spectroscopy Methods 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 235000002594 Solanum nigrum Nutrition 0.000 description 1
- 244000061457 Solanum nigrum Species 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 150000001336 alkenes Chemical class 0.000 description 1
- 125000000304 alkynyl group Chemical group 0.000 description 1
- 150000001540 azides Chemical class 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000005842 biochemical reaction Methods 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 229910001431 copper ion Inorganic materials 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 235000013601 eggs Nutrition 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 125000004185 ester group Chemical group 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 239000003068 molecular probe Substances 0.000 description 1
- JRZJOMJEPLMPRA-UHFFFAOYSA-N olefin Natural products CCCCCCCC=C JRZJOMJEPLMPRA-UHFFFAOYSA-N 0.000 description 1
- 230000010355 oscillation Effects 0.000 description 1
- 125000002467 phosphate group Chemical group [H]OP(=O)(O[H])O[*] 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 235000015497 potassium bicarbonate Nutrition 0.000 description 1
- 229910000028 potassium bicarbonate Inorganic materials 0.000 description 1
- 239000011736 potassium bicarbonate Substances 0.000 description 1
- 239000001103 potassium chloride Substances 0.000 description 1
- 235000011164 potassium chloride Nutrition 0.000 description 1
- TYJJADVDDVDEDZ-UHFFFAOYSA-M potassium hydrogencarbonate Chemical compound [K+].OC([O-])=O TYJJADVDDVDEDZ-UHFFFAOYSA-M 0.000 description 1
- 229940086066 potassium hydrogencarbonate Drugs 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 230000004224 protection Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 239000000376 reactant Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 238000010183 spectrum analysis Methods 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- GFISHBQNVWAVFU-UHFFFAOYSA-K terbium(iii) chloride Chemical compound Cl[Tb](Cl)Cl GFISHBQNVWAVFU-UHFFFAOYSA-K 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000029812 viral genome replication Effects 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
- C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
- C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
- A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N21/6456—Spatial resolved fluorescence measurements; Imaging
- G01N21/6458—Fluorescence microscopy
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1003—Carbocyclic compounds
- C09K2211/1007—Non-condensed systems
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1029—Heterocyclic compounds characterised by ligands containing one nitrogen atom as the heteroatom
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1074—Heterocyclic compounds characterised by ligands containing more than three nitrogen atoms as heteroatoms
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/18—Metal complexes
- C09K2211/182—Metal complexes of the rare earth metals, i.e. Sc, Y or lanthanide
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Pathology (AREA)
- General Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Analytical Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Materials Engineering (AREA)
- Biomedical Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Optics & Photonics (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
本发明公开一种生物正交激活的时间分辨响应型稀土探针及其制备方法与应用,属于生物标记及成像技术领域。本发明提供的稀土探针内含有四嗪基团,与Tb3+的特征荧光发生共振能量转移,致使荧光微弱;但稀土探针与D‑甘露糖环辛烯发生生物正交反应,进而探针内的荧光共振能量转移不能够发生,致使稀土离子Tb3+的特征荧光释放出来,同时探针溶液伴有从粉红色到无色的裸眼可见颜色变化,可用于肿瘤细胞的细胞膜成像;另外,该稀土探针具有良好的水溶性,将在解决对肿瘤细胞、斑马鱼等成像过程中所引起的自发荧光问题具有重要意义。
Description
技术领域
本发明属于生物标记及成像技术领域,特别涉及基于生物正交反应且具有时间分辨荧光响应性探针及其制备方法,具体涉及生物正交激活的时间分辨响应型稀土探针及其制备方法与应用。
背景技术
生物正交反应是一类能够在活体细胞内不干扰生物自身生化反应条件下进行的化学反应,主要包括有含有叠氮与炔基或磷酸基团、四嗪与烯烃或者反式环辛烯等,具有选择性高、反应迅速等特点。因此,发展生物正交反应探针在拓宽生物应用方面是十分必要的。
目前,该反应结合有机荧光小分子探针及其成像技术已广泛应用于生物标记及其成像领域。其中,对于糖蛋白的标记和成像在肿瘤细胞转移的基础研究、生物医学诊断方面具有重要意义。细胞膜表面的糖蛋白(Glycans)、糖磷脂作为细胞膜重要组成部分,在许多生理过程中扮演着非常重要的角色,如癌症细胞的异常增殖与转移、细胞生长、免疫保护、病毒的复制、炎症的产生等。
目前,对于糖蛋白的标记和成像方法主要是有机荧光小分子探针及纳米荧光材料等。【参见:a)J.R.Ha,L.Hao,G.Venkateswaran,Y.H.Huang,E.Garcia,S.Persad,Exp.CellRes.2014,321,153–166;b)N.Khidekel,S.B.Ficarro,E.C.Peters,L.C.Hsieh Wilson,Proc.Natl.Acad.Sci.USA 2004,101,13132–13137;c)W.Morelle,J.C.Michalski,Nat.Protoc.2007,2,1585–1602.】此类方法的主要缺点是破坏了糖蛋白的结构。近些年,以Diels-Alder反应为主的生物正交反应荧光探针在分子识别和医学诊断等领域受到广泛的关注,因为凭借该反应具有操作简单、反应物易于合成、特异性高等优点。【a)O.T.Keppler,R.Horstkorte,M.Pawlita,C.Schmidt,W.Reutter,Glycobiology 2001,11,11R–18R;b)D.H.Dube,C.R.Bertozzi,Curr.Opin.Chem.Biol.2003,7,616–625;c)T.-L.Hsu,S.R.Hanson,K.Kishikawa,S.K.Wang,M.Sawa,C.H.Wong,Proc.Natl.Acad.Sci.U.S.A.2007,104,2614–2619;d)J.Du,M.A.Meledeo,Z.Wang,H.S.Khanna,V.D.Paruchuri,K.J.Yarema,Glycobiology 2009,19,1382–1401;e)S.Stairs,A.A.Neves,H.Y.A.Wainman,H.Ireland-Zecchini,K.M.Brindle,F.J.Leeper,ChemBioChem 2013,14,1063–1067.】但此类探针主要缺点是:1)铜离子的毒性较大;2)荧光背景大,特别是生物细胞和组织的自发荧光,不利于大规模推广。因此,开发具有低毒、时间分辨特性的荧光探针在实际应用中仍然具有重要的意义。
发明内容
为了克服现有技术的缺点与不足,本发明的首要目的在于提供生物正交激活的时间分辨响应型稀土探针。该稀土探针以快速准确、灵敏无毒的实现对肿瘤细胞的细胞膜成像。
本发明的另一目的在于提供上述生物正交激活的时间分辨响应型稀土探针的制备方法。其制备是以7-氨基-2-酮基-4-喹啉酸为原料,通过偶联等过程得到含有四嗪官能团的中间体,此中间体进一步与二乙基三胺五乙酸、六水氯化铽进行螯合获得生物正交激活的时间分辨响应型稀土探针。
本发明的再一目的在于提供上述生物正交激活的时间分辨响应型稀土探针的应用。
本发明的目的通过下述技术方案实现:
生物正交激活的时间分辨响应型稀土探针,其结构式如式Ⅰ所示:
上述生物正交激活的时间分辨响应型稀土探针的制备方法,包括如下步骤:
(1)IX1中间体的制备:对7-氨基-2-酮基-4-喹啉酸进行羧基活化,活化的7-氨基-2-酮基-4-喹啉酸与4-(6-甲基-1,2,4,5-四嗪-3-基)苄胺在N,N-二甲基甲酰胺中室温搅拌,除去溶剂并用硅胶色谱分离得含有四嗪官能团的红色固体;
(2)IX2中间体的制备:对7-氨基-2-酮基-4-喹啉酸进行羧基活化,活化的7-氨基-2-酮基-4-喹啉酸与乙二胺反应的产物和4-(6-甲基-1,2,4,5-四嗪-3-基)苄胺与丁二酸酐反应的产物在N,N-二甲基甲酰胺中室温搅拌,除去溶剂,硅胶色谱分离得到含有四嗪官能团的红色固体;
(3)IX1或IX2制备:将步骤(1)或(2)得到的含有四嗪官能团的红色固体与二乙基三胺五乙酸在二甲基亚砜中搅拌,用乙醚沉淀干燥,再在二甲基亚砜/水(体积比,1:1)中与六水氯化铽搅拌反应,乙醚沉淀干燥得稀土探针IX1或稀土探针IX2,即生物正交激活的时间分辨响应型稀土探针。
步骤(1)、(2)中所述的对7-氨基-2-酮基-4-喹啉酸进行羧基活化的活化剂为1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐和N-羟基丁二酰亚胺;
所述的1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐、N-羟基丁二酰亚胺、7-氨基-2-酮基-4-喹啉酸的摩尔比为5~10:5~10:1;优选为5:5:1;
优选的,步骤(1)中所述的活化的7-氨基-2-酮基-4-喹啉酸与4-(6-甲基-1,2,4,5-四嗪-3-基)苄胺的摩尔比为2:2~5;进一步为1:1;
优选的,步骤(2)中所述的活化的7-氨基-2-酮基-4-喹啉酸与乙二胺反应的产物和4-(6-甲基-1,2,4,5-四嗪-3-基)苄胺与丁二酸酐反应的产物的摩尔比为1:1~5;进一步为1:1;
优选的,步骤(2)中所述的活化的7-氨基-2-酮基-4-喹啉酸与乙二胺的摩尔比为1:1~4;进一步为1:1;
优选的,步骤(2)中所述的4-(6-甲基-1,2,4,5-四嗪-3-基)苄胺与丁二酸酐的摩尔比为1:1~5;进一步为1:1;
优选的,步骤(3)中所述的含有四嗪官能团的红色固体、二乙基三胺五乙酸、六水氯化铽的摩尔比为1:1~2:1~2;进一步为1:1:1;
优选的,步骤(1)、(2)、(3)中所述的搅拌的时间均为6~48小时;进一步为12小时。
上述生物正交激活的时间分辨响应型稀土探针在细胞膜成像中的应用。
优选的,所述的生物正交激活的时间分辨响应型稀土探针在肿瘤细胞的细胞膜成像或斑马鱼成像中的应用。
进一步优选的,所述的肿瘤细胞为肺癌细胞。
本发明相对于现有技术具有如下的优点及效果:
本发明提供的生物正交激活的时间分辨响应型稀土探针,具体为式Ⅰ所示的结构式,可用于肿瘤细胞的细胞膜成像。此稀土探针内含有四嗪基团,与Tb3+的特征荧光发生共振能量转移,致使荧光微弱;但稀土探针与D-甘露糖环辛烯发生生物正交反应,进而探针内的荧光共振能量转移不能够发生,致使稀土离子Tb3+的特征荧光释放出来,同时探针溶液伴有从粉红色到无色的裸眼可见颜色变化;另外,该稀土探针具有良好的水溶性,将在解决对肿瘤细胞、斑马鱼等成像过程中所引起的自发荧光问题具有重要意义。
附图说明
图1是实施例1、2、3中生物正交激活的时间分辨响应型稀土探针的制备合成路线。
图2是实施例4中生物正交激活的时间分辨响应型稀土探针的光谱分析;其中,a)为IX1、IX1与TCO反应前后的紫外吸收图;c)为IX2、IX2与TCO反应前后的紫外吸收图;b)为IX1、IX1与TCO反应前后的荧光光谱图,5D4-7F6、5D4-7F5、5D4-7F4、5D4-7F3等表示稀土离子Tb3+的特征能级跃迁,分别对应波长为490nm、545nm、585nm、622nm的发射峰;d)为IX2、IX2与TCO反应前后的荧光光谱图。
图3是实施例5中生物正交激活的时间分辨响应型稀土探针反应前后的荧光寿命的变化。
图4是实施例1中所述稀土探针IX1的中间体S1核磁共振氢谱图。
图5是实施例2中所述稀土探针IX1的中间体S2的电感耦合等离子体质谱(ICP-MS)图。
图6是实施例2中所述稀土探针IX1的电感耦合等离子体质谱图。
图7是实施例3中所述稀土探针IX2的中间体S3的电感耦合等离子体质谱图。
图8是实施例3中所述稀土探针IX2的中间体S4的电感耦合等离子体质谱图。
图9是实施例3中所述稀土探针IX2的电感耦合等离子体质谱图。
图10是实施例6中所述生物正交激活的时间分辨响应型稀土探针IX1对肿瘤细胞的细胞膜成像,左和右图分别为孵育含有TCO甘露糖(Ac4ManNTCO)和不含TCO的甘露糖(Ac4ManNH2)的细胞中细胞激光共聚焦成像图。
图11是实施例7中所述生物正交激活的时间分辨响应型稀土探针IX1对斑马鱼进行成像,上和下图分别为孵育含有TCO甘露糖(Ac4ManNTCO)和不含TCO的甘露糖(Ac4ManNH2)的斑马鱼激光共聚焦成像图。
具体实施方式
下面结合实施例及附图对本发明作进一步详细的描述,但本发明的实施方式不限于此。
下列实施例中未注明具体实验条件的实验方法,通常按照常规实验条件或按照制造厂商所建议的实验条件。
实施例中的7-氨基-2-酮基-4-喹啉酸需参考文献:Bioconjugate Chem.2004,15,1088-1094;Bioconjugate Chem.2011,22,1402-1409;4-(6-甲基-1,2,4,5-四嗪-3-基)苄胺(CAS号:1345955-28-3)参考文献:Angew.Chem.Int.Ed.2009,48,7013-7016;Bioconjugate Chem.2011,22,2263-2270。
实施例1:
(1)本发明所提供的生物正交激活的时间分辨响应型稀土探针其合成步骤分别如图1所示,具体如下:IX1中间体S1的制备:将7-氨基-2-酮基-4-喹啉酸(43.6mg,0.2mmol)与N-羟基丁二酰亚胺NHS(115.0mg,1.0mmol)、1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐EDC·HCl(190.8mg,1.0mmol)溶于10mL N,N-二甲基甲酰胺并在室温下搅拌过夜进行羧基活化,结束后用饱和食盐水沉淀,离心干燥得黄褐色固体(即活化的7-氨基-2-酮基-4-喹啉酸);继续将黄褐色固体与4-(6-甲基-1,2,4,5-四嗪-3-基)苄胺(40.2mg,0.2mmol)在N,N-二甲基甲酰胺中室温搅拌过夜,除去溶剂,硅胶色谱分离得红色固体S1(66.8mg),产率83.3%。
表征数据(图4):1HNMR(500M,d6-DMSO):δ=2.99(s,2H),3.16(d,J=5Hz,1H),4.40(d,J=5.0Hz,2H),5.74(d,J=15.0Hz,2H),6.05(s,1H),6.38(s,1H),6.44(d,J=10.0Hz,1H),7.42(d,J=10.0Hz,1H),7.49(d,J=10.0Hz,2H),8.38(d,J=15.0Hz,2H),8.74(t,J=5.0Hz,1H),11.23(s,1H)。
红色固体S1的结构式如下所示:
实施例2:
(1)IX1中间体S2的制备:在室温下,去取实施例1得到的红色固体S1(20.5mg,0.05mmol)、二乙基三胺五乙酸DTPA(19.7mg,0.05mmol)溶于3mL二甲基亚砜中,搅拌过夜反应,然后用乙醚沉淀干燥得固体S2(34.3mg),产率88.4%。
表征数据:ICP-MS(图5):calcd.for[M+]776.76,found:713.40.
(2)IX1的制备:在室温下,去取步骤(1)得到的固体S2(7.8mg,0.01mmol)、六水氯化铽TbCl3·6H2O(3.8mg,0.01mmol)溶于1mL二甲基亚砜/水(体积比,1:1)中,搅拌过夜反应,然后用乙醚沉淀干燥得稀土探针IX1(8.41mg),产率90.1%。
表征数据:ICP-MS(图6):calcd.for[M+]931.66,found:931.20.
实施例3:
(1)IX2中间体S3、S4的制备:取实施例1中活化的7-氨基-2-酮基-4-喹啉酸(33.2mg,0.1mmol)溶于含有乙二胺(6mg,0.1mmol)的10mL N,N-二甲基甲酰胺中,在室温下搅拌过夜,用TCL监测反应完成后,将4-(6-甲基-1,2,4,5-四嗪-3-基)苄胺(30.1mg,0.1mmol)与丁二酸酐(10.0mg,0.1mmol)反应的产物在N,N-二甲基甲酰胺中室温搅拌过夜,除去溶剂,硅胶色谱分离得红色固体S3。在室温下,再将红色固体S3、二乙基三胺五乙酸(39.5mg,0.1mmol)溶于4mL二甲基亚砜中,搅拌过夜反应,然后用乙醚沉淀干燥得其固体S4(68.3mg),产率74.4%。
S3表征数据:ICP-MS(图7):calcd.for[M+]526.59,found:525.10.
红色固体S3的结构式如下所示:
S4表征数据:ICP-MS(图8):calcd.for[M+]918.92,found:917.30.
(2)IX2的制备:在室温下,去取步骤(1)得到的固体S4(9.2mg,0.01mmol)、六水氯化铽(3.8mg,0.01mmol)溶于1mL二甲基亚砜/水(体积比,1:1)中,搅拌过夜反应,然后用乙醚沉淀干燥得稀土探针IX2(9.33mg),产率86.8%。
IX2表征数据:ICP-MS(图9):calcd.for[M+]1073.8,found:1073.20.
实施例4:
对实施例2、3得到的稀土探针在水与甲醇混合介质中与环辛烯(TCO)反应的紫外吸收和荧光强度、荧光寿命变化分析:配置浓度为10.0μmol/L的稀土探针,再分别加入20.0μmol/L环辛烯于其中振荡反应1分钟,在测试记录该稀土探针的吸收光谱(图2a与2c)和荧光光谱(图2b与2d)的变化。通过分析吸收光谱和荧光光谱强度的变化,说明两种稀土探针具有荧光响应。同时,通过分析荧光光谱强度的变化,建议孵育细胞的探针浓度为50μmol/L。
实施例5:
对实施例2、3得到的稀土探针IX1、IX2与环辛烯反应前后荧光寿命的测试:首先,分别用时间分辨荧光分析仪测试10μmol/L的稀土探针IX1、IX2的荧光寿命,再加入20μmol/L的环辛烯,振荡反应1分钟,然后测其各自反应后的荧光寿命,其结果见图3。通过分析可知两种探针在反应后的荧光寿命都大大延长,具有实现时间分辨成像的潜能。
实施例6:
对实施例2得到的稀土探针IX1对肿瘤细胞A549(市售细胞)的细胞膜进行成像分析:首先,将50μmol/L的D-甘露糖环辛烯(Ac4ManNTCO)和50μmol/L的D-甘露糖(Ac4ManNH2)分别加入到细胞培养基DMEM中,37℃下孵育48小时,吸出培养液,并用PBS缓冲溶液冲洗三次,再加入新的DMEM培养液,两份均加入50μmol/L的稀土探针IX1,37℃下孵育8小时后,分别进行激光共聚焦显微成像,如图10,从图中可以看出,该稀土探针可以对细胞膜进行成像,获得良好的细胞成像图。图10左边为D-甘露糖环辛烯孵育的细胞双光子激光共聚焦成像图,右边为D-甘露糖孵育的细胞激光共聚焦成像图。
实施例7:
对实施例2得到的稀土探针IX1对斑马鱼进行成像分析:首先,将5mmol/L的D-甘露糖环辛烯和5mmol/L的D-甘露糖分别通过卵内注射到斑马鱼受精卵内,37℃下孵育48小时,吸出培养液,再加入新的斑马鱼培养液(氯化钠,150mmol/L;氯化钾,0.5mmol/L;氯化钙,1.0mmol/L;磷酸二氢钾,0.37mmol/L;磷酸一氢钾,0.05mmol/L;硫酸镁,2.0mmol/L;碳酸氢钾,0.71mmol/L的去离子水溶液,pH为7.4),两份均加入50μmol/L的稀土探针IX1,37℃下孵育8小时后,分别进行双光子激光共聚焦显微成像,如图11,从图中可以看出,该稀土探针可以对斑马鱼进行成像,获得良好的成像图。图11上排图为D-甘露糖环辛烯孵育的斑马鱼激光共聚焦成像图,图11下排图为D-甘露糖孵育的斑马鱼激光共聚焦成像图。
上述实施例为本发明较佳的实施方式,但本发明的实施方式并不受上述实施例的限制,其他的任何未背离本发明的精神实质与原理下所作的改变、修饰、替代、组合、简化,均应为等效的置换方式,都包含在本发明的保护范围之内。
Claims (9)
2.权利要求1所述的生物正交激活的时间分辨响应型稀土探针的制备方法,其特征在于包括如下步骤:
(1)IX1中间体的制备:对7-氨基-2-酮基-4-喹啉酸进行羧基活化,活化的7-氨基-2-酮基-4-喹啉酸与4-(6-甲基-1,2,4,5-四嗪-3-基)苄胺在N,N-二甲基甲酰胺中室温搅拌,除去溶剂并用硅胶色谱分离得含有四嗪官能团的红色固体;
(2)IX2中间体的制备:对7-氨基-2-酮基-4-喹啉酸进行羧基活化,活化的7-氨基-2-酮基-4-喹啉酸与乙二胺反应的产物和4-(6-甲基-1,2,4,5-四嗪-3-基)苄胺与丁二酸酐反应的产物在N,N-二甲基甲酰胺中室温搅拌,除去溶剂,硅胶色谱分离得到含有四嗪官能团的红色固体;
(3)IX1或IX2制备:将步骤(1)或(2)得到的含有四嗪官能团的红色固体与二乙基三胺五乙酸在二甲基亚砜中搅拌,用乙醚沉淀干燥,再在体积比1:1的二甲基亚砜/水中与六水氯化铽搅拌反应,乙醚沉淀干燥得稀土探针IX1或稀土探针IX2,即生物正交激活的时间分辨响应型稀土探针。
3.根据权利要求2所述的制备方法,其特征在于:
步骤(1)、(2)中所述的对7-氨基-2-酮基-4-喹啉酸进行羧基活化的活化剂为1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐和N-羟基丁二酰亚胺;
所述的1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐、N-羟基丁二酰亚胺、7-氨基-2-酮基-4-喹啉酸的摩尔比为5~10:5~10:1;
步骤(1)、(2)、(3)中所述的搅拌的时间均为6~48小时。
4.根据权利要求3所述的制备方法,其特征在于:
所述的1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐、N-羟基丁二酰亚胺、7-氨基-2-酮基-4-喹啉酸的摩尔比为5:5:1;
步骤(1)、(2)、(3)中所述的搅拌的时间均为12小时。
5.根据权利要求2~4任一项所述的制备方法,其特征在于:
步骤(1)中所述的活化的7-氨基-2-酮基-4-喹啉酸与4-(6-甲基-1,2,4,5-四嗪-3-基)苄胺的摩尔比为2:2~5;
步骤(2)中所述的活化的7-氨基-2-酮基-4-喹啉酸与乙二胺反应的产物和4-(6-甲基-1,2,4,5-四嗪-3-基)苄胺与丁二酸酐反应的产物的摩尔比为1:1~5;
步骤(2)中所述的活化的7-氨基-2-酮基-4-喹啉酸与乙二胺的摩尔比为1:1~4;
步骤(2)中所述的4-(6-甲基-1,2,4,5-四嗪-3-基)苄胺与丁二酸酐的摩尔比为1:1~5;
步骤(3)中所述的含有四嗪官能团的红色固体、二乙基三胺五乙酸、六水氯化铽的摩尔比为1:1~2:1~2。
6.根据权利要求5所述的制备方法,其特征在于:
步骤(1)中所述的活化的7-氨基-2-酮基-4-喹啉酸与4-(6-甲基-1,2,4,5-四嗪-3-基)苄胺的摩尔比为1:1;
步骤(2)中所述的活化的7-氨基-2-酮基-4-喹啉酸与乙二胺反应的产物和4-(6-甲基-1,2,4,5-四嗪-3-基)苄胺与丁二酸酐反应的产物的摩尔比为1:1;
步骤(2)中所述的活化的7-氨基-2-酮基-4-喹啉酸与乙二胺的摩尔比为1:1;
步骤(2)中所述的4-(6-甲基-1,2,4,5-四嗪-3-基)苄胺与丁二酸酐的摩尔比为1:1;
步骤(3)中所述的含有四嗪官能团的红色固体、二乙基三胺五乙酸、六水氯化铽的摩尔比为1:1:1。
7.权利要求1所述的生物正交激活的时间分辨响应型稀土探针在制备细胞膜成像的试剂中的应用。
8.根据权利要求7所述的应用,其特征在于:
所述的生物正交激活的时间分辨响应型稀土探针在制备肿瘤细胞的细胞膜成像的试剂或制备斑马鱼成像的试剂中的应用。
9.根据权利要求8所述的应用,其特征在于:
所述的肿瘤细胞为肺癌细胞。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201911266381.7A CN111732576B (zh) | 2019-12-11 | 2019-12-11 | 生物正交激活的时间分辨响应型稀土探针及其制备方法与应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201911266381.7A CN111732576B (zh) | 2019-12-11 | 2019-12-11 | 生物正交激活的时间分辨响应型稀土探针及其制备方法与应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN111732576A CN111732576A (zh) | 2020-10-02 |
CN111732576B true CN111732576B (zh) | 2021-08-06 |
Family
ID=72646091
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201911266381.7A Expired - Fee Related CN111732576B (zh) | 2019-12-11 | 2019-12-11 | 生物正交激活的时间分辨响应型稀土探针及其制备方法与应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN111732576B (zh) |
Family Cites Families (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP1623979A4 (en) * | 2003-02-24 | 2008-11-26 | Japan Science & Tech Agency | FLUORESCENT LANTHANIDE COMPLEX |
JP4951759B2 (ja) * | 2007-01-01 | 2012-06-13 | 国立大学法人九州工業大学 | レドックス応答性発光プローブ及びそれを用いる検出方法 |
WO2012121746A2 (en) * | 2011-03-09 | 2012-09-13 | The General Hospital Corporation | Imaging beta cell mass |
-
2019
- 2019-12-11 CN CN201911266381.7A patent/CN111732576B/zh not_active Expired - Fee Related
Also Published As
Publication number | Publication date |
---|---|
CN111732576A (zh) | 2020-10-02 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN108514647B (zh) | 基质金属蛋白酶-2特异性多模态分子影像探针及其制备方法与在制备肿瘤成像剂中的应用 | |
CN107325095B (zh) | 一种溶酶体次氯酸荧光探针及其制备方法和应用 | |
CN112409322B (zh) | Ggt激活型化学发光探针及其合成方法和应用 | |
CN105924394A (zh) | 一种双光子甲醛荧光探针及其制备与应用 | |
CN106892947B (zh) | 一种含有(肼基羰基)二茂铁配体的铱配合物及其制备方法和应用 | |
CN109438319B (zh) | 一种检测亮氨酸氨肽酶的化合物及其制备方法和应用 | |
CN109336815B (zh) | 一种检测细胞内质网内次氯酸的双光子荧光探针 | |
CN106243170B (zh) | 具有聚集诱导荧光增强特性的β-半乳糖苷酶传感器的合成及应用 | |
CN110272734A (zh) | 一种用于no检测的高量子产率碳量子点制备方法及其应用 | |
CN109336835B (zh) | 用于检测髓过氧化物酶活性荧光探针及其制备方法和应用 | |
CN114478473B (zh) | 一种亮氨酸氨基肽酶化学发光检测试剂的合成及应用 | |
CN112062755B (zh) | 用于检测天冬氨酰氨肽酶的近红外荧光分子探针、制备方法及用途 | |
CN110078665A (zh) | 一种内质网靶向的检测次氯酸的荧光探针和应用 | |
CN112500386B (zh) | 基于吡罗红肟的近红外HClO荧光探针、制备及其应用 | |
Zhang et al. | A lysosome-targetable fluorescent probe for the simultaneous sensing of Cys/Hcy and GSH from different emission channels | |
CN116178349A (zh) | 一种检测半胱氨酸的高尔基体靶向近红外荧光探针、其制备方法和应用 | |
CN112266351A (zh) | 一种双光子比率荧光探针及其制备方法与应用 | |
Tanaka et al. | Ratiometric multimodal chemosensors based on cubic silsesquioxanes for monitoring solvent polarity | |
CN108752275B (zh) | 一种pH荧光探针及其制备方法和应用 | |
CN111732576B (zh) | 生物正交激活的时间分辨响应型稀土探针及其制备方法与应用 | |
US8187825B2 (en) | Thiol detection method | |
CN114957299B (zh) | 用于检测凋亡细胞的荧光探针及其制备方法 | |
EP2758391B1 (fr) | Sondes luminescentes pour le marquage biologique et l'imagerie, et leur procédé de préparation. | |
JP6675125B2 (ja) | pH依存性蛍光化合物 | |
CN110687087B (zh) | 一种溶酶体三磷酸腺苷识别碳点的制备方法及应用 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant | ||
CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20210806 |
|
CF01 | Termination of patent right due to non-payment of annual fee |